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为了研究铅(Pb)和邻苯二甲酸二丁酯(DBP)单独及联合暴露对斑马鱼胚胎神经发育和细胞凋亡相关基因表达的影响,考察两者对斑马鱼胚胎神经系统的毒性作用。将720个斑马鱼胚胎按3×3析因设计随机分为9组,以Pb(0、0.01 mg/L和1 mg/L)和DBP(0、0.005 mg/L和0.5 mg/L)单独及联合对斑马鱼胚胎暴露120 h。观察、记录胚胎的孵化、死亡情况,采用荧光定量PCR测定斑马鱼胚胎神经发育相关基因(NR1A、NR2A、NR2D)和神经细胞凋亡相关基因(bcl-2、c-fos)的表达变化情况。与空白对照组相比,各暴露组均可致斑马鱼胚胎孵化率降低、死亡率增高(P<0.05);联合暴露组NR1A、NR2A、NR2D、c-fos基因表达水平显著升高,bcl-2基因表达水平显著下降(P<0.05)。Pb和DBP均可影响斑马鱼胚胎的发育,且具有一定的神经毒性,而二者联合暴露毒性复杂,对斑马鱼胚胎孵化率和死亡率的影响有交互作用,对神经发育和细胞凋亡相关基因无交互作用,其机制有待进一步研究。
To investigate the effects of Pb and DBP exposure on the neural development and apoptosis-related gene expression in zebrafish embryos, the toxic effects of both on the nervous system of zebrafish embryos were investigated. 720 zebrafish embryos were randomly divided into 9 groups according to 3 × 3 factorial design. Pb (0,0.01 mg / L and 1 mg / L) and DBP (0,0.005 mg / L and 0.5 mg / L) alone And combined exposure to zebrafish embryos 120 h. The hatching and death of embryos were observed and recorded. Fluorescent quantitative PCR was used to detect the expression of neuronal development related genes (NR1A, NR2A, NR2D) and neuronal apoptosis-related genes (bcl-2, c-fos) in zebrafish embryos. Compared with the blank control group, the hatching rate and the mortality of zebrafish embryos in each exposure group were significantly decreased (P <0.05). The expression of NR1A, NR2A, NR2D and c-fos genes in the co-exposure group was significantly increased 2 gene expression decreased significantly (P <0.05). Both Pb and DBP can affect the development of zebrafish embryos and have certain neurotoxicity. However, the combined exposure toxicity of Pb and DBP has an interaction effect on zebrafish embryo hatching rate and mortality, and is related to neurodevelopment and apoptosis Gene no interaction, the mechanism needs further study.